Vectorial and nonvectorial transphosphorylation catalyzed by enzymes II of the bacterial phosphotransferase system.
Vectorial transphosphorylation of hexitols, catalyzed by enzymes II of the bacterial phosphotransferase system, was studied in intact cells and membrane vesicles of Escherichia coli. In strains depleted of phosphoenolpyruvate and unable to metabolize the internal hexitol phosphate, internal mannitol-1-phosphate stimulated uptake of extracellular [14C]mannitol, whereas external mannitol stimulated release of [14C]mannitol from the intracellular [14C]mannitol-1-phosphate pool. The stoichiometry of mannitol uptake to mannitol release was 1:1. Glucitol did not promote release of [14C]mannitol from the mannitol phosphate pool but stimulated release of [14C]glucitol from internal glucitol phosphate pools when the glucitol enzyme II was induced to high levels. In E coli cells and membrane vesicles, both vectorial and nonvectorial transphosphorylation reactions of hexitols and hexoses were demonstrated. The nonvectorial reactions, but not the vectorial reactions, catalyzed by the mannitol and glucose enzymes II, were inhibited by p-chloromercuriphenyl sulfonate, a membrane-impermeable sulfhydryl reagent which inactivates enzymes II. Similarly, glucose-6-sulfate, an inhibitor of the glucose enzyme II-catalyzed transphosphorylation reaction, specifically inhibited the nonvectorial reaction. This compound was shown to be a noncompetitive inhibitor of methyl alpha-glucoside phosphorylation employing phospho-HPr as the phosphate donor. It apparently exerts its inhibitory effect by exclusive binding to the sugar phosphate binding site on the enzyme II complex. The results are consistent with the conclusion that enzymes II can exist in two distinct dispositions in the membrane, one of which catalyzes vectorial transphosphorylation, and the other catalyzes nonvectorial transphosphorylation.
D002462:Cell Membrane / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology*; D008353:Mannitol / Q000378:metabolism*; D008757:Methylglucosides / Q000378:metabolism; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000378:metabolism*; D010766:Phosphorylation; D013012:Sorbitol / Q000378:metabolism*
D008757:Methylglucosides; D008353:Mannitol; D013012:Sorbitol; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System